摘要
背景:目前作为骨组织工程的种子细胞主要是骨髓间充质干细胞,但其在骨髓中的含量极低,只有0.001%-0.01%,同时分离纯化难度大。目的:探讨人胎盘来源间充质干细胞的生物学性能及与三维多孔羟基磷灰石陶瓷支架的生物相容性。方法:人胎盘来源间充质干细胞进行形态学观察和流式细胞仪鉴定;将人胎盘来源间充质干细胞经成骨、成脂、成软骨诱导分化培养3周,进行茜素红S、油红O、甲苯胺蓝染色,鉴定其多向分化潜能。将人胎盘来源间充质干细胞接种于三维多孔羟基磷灰石陶瓷支架,通过DAPI染色、扫描电镜观察人胎盘来源间充质干细胞在支架表面的黏附情况。结果与结论:(1)人胎盘来源间充质干细胞呈长梭形,大小均匀一致;(2)人胎盘来源间充质干细胞高表达CD29,CD90等干细胞表面标志,不表达CD45,CD106等造血细胞表面标志;(3)诱导培养3周后,茜素红S染色可以看到红色的矿化结节,油红O染色观察到红色的脂滴,甲苯胺蓝染色可以看到软骨细胞染成蓝色;(4)DAPI染色、扫描电镜均观察到人胎盘来源间充质干细胞在支架表面能很好地黏附,可以作为组织工程良好的种子细胞。
BACKGROUND： At present bone marrow mesenchymal stem cells act as the main seed cells in bone tissue engineering, but only 0.001%-0.01% cells are in the bone with difficulty in cell separation and purification. OBJECTIVE： To explore the biological characterization of human placenta derived mesenchymal stem cells and biocompatibility with three-dimensional porous hydroxyapatite ceramic scaffold.METHODS： Human placenta derived mesenchymal stem cells were morphologically observed and identified usingflow cytometry, followed by osteogenic, adipogenic, chondrogenic induction for 3 weeks. Afterwards, the potential of multi-directional differentiation was identified by alizarin red S, oil red O and toluidine blue staining. DAPI staining was used to observe the adhesion of cells on the surface of the hydroxyapatite ceramic scaffold under scanning electron microscope. RESULTS AND CONCLUSION： The human placenta derived mesenchymal stem cells showed long spindle shape and uniform size under the microscope; they highly expressed CD29 and D90, but did not express CD45 and CD106. Following induction, mineralized nodules were observed by alizarin red S staining, lipid droplets by oil red O staining and blue-dyed toluidine blue staining. These cells adhered well to the scaffold surface, indicting they are suitable for bone tissue engineering.
作者
伍佳
温永梅
吕欣荣
牟雁东
Wu Jia Wen Yong-mei Lv Xin-rong Mu Yan-dong(Southwest Medical University, Luzhou 646000, Sichuan Province, China Sichuan Provincial People＇s Hospital, Chengdu 610000, Sichuan Province, China)
出处
《中国组织工程研究》
CAS
北大核心
2017年第5期755-759,共5页
Chinese Journal of Tissue Engineering Research
基金
四川省科技厅项目(2016TD0008
2014JY0187)~~
